title: Characterization and functional analysis of the 5' flanking region of Sparus aurata myostatin-1 gene.
abstract: Myostatin (MSTN) is a member of the transforming growth factor-beta superfamily that functions as a negative regulator of skeletal muscle development and growth in mammals. Although several MSTN promoters were described in fish, no functional analysis was reported so far. Here, the 5' flanking region (1372 bp) of the MSTN-1 gene of the marine fish Sparus aurata (saMSTN-1) was cloned, sequenced and characterized. It contains two consensus sequences for TATA box (TATAA), a CAAT box, ten putative E-boxes known as binding sites to myogenic basic helix-loop-helix transcription factors (TFs) and two putative binding sites to TF Myocyte enhancing factor-2 (MEF2). In addition, it has several putative binding sites to TF Pit-1a and several response elements to nuclear receptors (GRE, ERE, PRE, ARE, TRE, RARE and PPARRE) and cAMP-response elements. Transcriptional activity of five genomic fragments (truncated at their upstream region) of 372, 941, 972, 1113 and 1355 bp was studied in vitro, using transient transfection in A204 cells. All constructs directed luciferase activity, with the highest activity obtained by the 1113 bp fragment. These experiments show that all five genomic fragments are functional MSTN promoters and differences in promoter activity might be due to presence of enhancers and/or repressor sequences, regulating MSTN promoter activity.
authors.affiliation: Department of Marine Biology and Biotechnology, Israel Oceanographic and Limnological Research, Tel-Shikmona, Haifa, Israel. bruria@ocean.org.il
authors.forename: Bruria
authors.initials: B
authors.lastname: Funkenstein
authors.forename: Viki
authors.initials: V
authors.lastname: Balas
authors.forename: Yanai
authors.initials: Y
authors.lastname: Rebhan
authors.forename: Anna
authors.initials: A
authors.lastname: Pliatner
mesh_terms: D024506:5' Flanking Region / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D017124:Conserved Sequence; D008969:Molecular Sequence Data; D055435:Myostatin / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D021541:Sea Bream / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D055435:Myostatin; D014157:Transcription Factors